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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
NeuroCult NS-A Proliferation Kit (Stem Cell Technologies cat# 5751)
20 μg/mL EGF: use 1 μL/1 mL culture medium. To prepare 20 μg/mL EGF stock solution, aseptically combine:
Then further dilute by adding 4 mL PBS/0.1% BSA
20 μg/mL β-FGF: use 1 μL/1 mL culture medium. To prepare 20 μg/mL EGF stock solution, aseptically combine:
Then further dilute by adding 4 mL PBS/0.1% BSA
Store in working aliquots at -20°C. EGF and β-FGF is stable for 1 year when prepared and stored as directed. Do not repeat freeze thaw.
2 mg/mL Heparin Solution (Stem Cell Technologies cat# 07980): use 1 μL/1 mL culture medium. Store solution at 4°C.
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Cusulin C, et al. Precursor States of Brain Tumor Initiating Cell Lines Are Predictive of Survival in Xenografts and Associated with Glioblastoma Subtypes. Stem Cell Reports 5(1): 1-9, 2015. PubMed: 26095605
Kelly JJ, et al. Proliferation of human glioblastoma stem cells occurs independently of exogenous mitogens. Stem Cells 27(8): 1722-33, 2009. PubMed: 19544433
Luchman A, et al. Dual mTORC1/2 Blockade Inhibits Glioblastoma Brain Tumor Intiating Cells In Vitro and In Vivo and Synergizes with Temozolomide to Increase Orthotopic Xenograft Survival. Clin Cancer Res 20(22): 5756-67, 2014. PubMed: 25316808
Stechishin O, et al. On-target JAK2/STAT3 inhibition slows disease progression in orthotopic xenografts of human glioblastoma brain tumor stem cells. Neuro Oncol 15(2): 198-207, 2013. PubMed: 23262510